Combination of analytical and microbiological techniques to study the antimicrobial activity of a new active food packaging containing cinnamon or oregano against E. coli and S. aureus.

The aim of this work is the optimization and application of a group of analytical and microbiological techniques in the study of the activity of essential oils (EOs) incorporated in a new antimicrobial packaging material and the research in depth of the interaction between the microbial cells and the individual compounds present in the active material. For this purpose the antimicrobial activity of the active packaging containing cinnamon or oregano was evaluated against E. coli and S. aureus. The vapour phase activity and the direct contact between the antimicrobial agents themselves, or once incorporated in the packaging material, and the microbial cells have been studied. The direct contact was studied using a broth dilution method. The vapour phase was evaluated by using a new method which involves the use of a filter disk containing the EOs. Furthermore, the kill time assay was used to determine the exposure time for the maximum efficiency in packaging, and transmission electron microscopy was used to investigate the antimicrobial activity and the possible mechanism of action against E. coli and S. aureus. Finally, the compounds absorbed by cells were identified. The results showed that the techniques used provide relevant information about the antibacterial activity of cinnamon and oregano in direct contact as well as in the vapour phase. The antimicrobial packaging showed a fast efficiency which supports its likely application as a food packaging material. Bacteria treated with EOs exhibit a wide range of significant abnormalities; these include formation of blebs, coagulation of cytoplasmatic constituents, collapse of the cell structure and lack of cytoplasmatic material. Some of these observations are correlated to the ability of some of these substances to disrupt envelop structure, especially the inner membrane. After an extraction from dead cells, cinnamaldehyde was detected by GC-MS in E. coli exposed to the active packaging containing cinnamon.